Molecular epidemiology of Hepatitis delta virus infection in Minas Gerais state from Brazil, an area outside the hyperendemic region of the Amazon Basin

BACKGROUND Hepatitis delta virus (HDV) infections in hepatitis B virus (HBV) carriers are the most severe form of viral hepatitis. HDV prevalence is high in the Brazilian Amazon, but studies in other regions of the country are still scarce and often underestimated its prevalence by including a small numbers of individuals. OBJECTIVE This study aimed to determine the serological prevalence of hepatitis D, the genotypes circulating and to evaluate the associated risk factors for acquisition of HDV in Minas Gerais state, Brazil. METHODS We screened plasma samples (n = 498) from HBV chronic carriers for anti-HD antibodies using a commercial enzyme-linked immunosorbent assay (ELISA) kit. For those samples that were positive for anti-HD antibodies, we performed a reverse transcriptase (RT) nested-polymerase chain reaction (nested-PCR) in order to detect the viral genome and identify the viral genotypes circulating in the state. FINDINGS The prevalence was 6.22% (31/498). Blood transfusion was the only risk factor associated with HDV infection [risk ratio: 3.73; 95% confidence interval (CI): 1.44 to 9.65]. For 26 anti-HD positive patients, HDAg gene sequences were determined and in all patients HDV genotype 1 was found. CONCLUSIONS This study confirmed the circulation of HDV in Minas Gerais, an area previously considered non-endemic for hepatitis D in Brazil. The prevalence found in this study is much higher when compared to other studies performed in Brazil, probably because the population in our study was selected with minimal bias. Furthermore, in 26 anti-HD positive plasma samples, we were also able to detect the viral genome, indicating that these patients were experienced an active infection at the time of sample collection. These findings emphasise the importance of anti-HD testing in HBV infected individuals, which may contribute to this disease control in Brazil.

Antidiarrheal activity of extracts from Maytenus gonoclada and inhibition of Dengue virus by lupeol

ABSTRACT Diarrhea is an infectious disease caused by bacterial, virus, or protozoan, and dengue is caused by virus, included among the neglected diseases in several underdeveloped and developing countries, with an urgent demand for new drugs. Considering the antidiarrheal potential of species of Maytenus genus, a phytochemical investigation followed by antibacterial activity test with extracts of branches and heartwood and bark of roots from Maytenus gonoclada were conducted. Moreover, due the frequency of isolation of lupeol from Maytenus genus the antiviral activity against Dengue virus and cytotoxicity of lupeol and its complex with β-cyclodextrins were also tested. The results indicated the bioactivity of ethyl acetate extract from branches and ethanol extract from heartwood of roots of M. gonoclada against diarrheagenic bacteria. The lupeol showed potent activity against Dengue virus and low cytotoxicity in LLC-MK2 cells, but its complex with β-cyclodextrin was inactive. Considering the importance of novel and selective antiviral drug candidates the results seem to be promising.

Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.

Antiviral activities of plants occurring in the state of Minas Gerais, Brazil: Part 2. Screening Bignoniaceae species / Atividade antiviral de extratos de plantas coletadas no estado de Minas Gerais: Parte 2. Triagem de Bignoniaceae

Ethanol extracts of eighteen Bignoniaceae species have been evaluated by the MTT assay for cytotoxicity in Vero cells and for antiviral activity against Human herpes virus type 1, Vaccinia virus and murine Encephalomyocarditis virus. Among such species, seven are reported to be of traditional medicinal use No cytotoxicity was observed for most of the extracts up to the concentration of 500 μg/mL. Fourteen (50 percent) of the 28 extracts assayed have disclosed antiviral activity with EC50 values in the range of 4.6+0.3 to 377.2+17.7 μg/mL. Only two species, Arrabidaea samydoides and Callichlamys latifolia, have shown activity against all the three viruses. The extracts were chemically characterized by their TLC and HPLC-DAD profiles. Mangiferin is the major constituent of A. samydoides but the isolated compound has been less active than the crude extract. This is the first report on the antiviral evaluation of the eighteen Bignoniaceae species assayed.

Extratos etanólicos de dezoito espécies vegetais pertencentes à família Bignoniaceae, das quais sete são descritas como de uso medicinal, foram avaliados, pelo ensaio colorimétrico do MTT, para atividades citotóxica, em células Vero, e antiviral, frente aos vírus herpes simplex-tipo 1, vaccinia e encefalomiocardite murina. A maior parte dos extratos não apresentou citotoxicidade até a concentração de 500 μg/mL. Dos 28 extratos testados quatorze (50 por cento) apresentaram atividade antiviral com valores de CE50 na faixa de 4,6+03 a 377,2+17,7 μg/mL. Somente duas espécies, Arrabidaea samydoides e Callichlamys latifolia, foram ativas frente aos três vírus. Os extratos foram caracterizados pelos seus perfís cromatográficos em CCD e CLAE-FR. Análises por CLAE-FR mostraram que a mangiferina é o constituinte majoritário em A. samydoides mas a substância isolada foi menos ativa do que o extrato bruto. Esta é a primeira vez que se relata a atividade antiviral de extratos das dezoito espécies avaliadas.

Vaccinia virus regulates expression of p21WAF1/Cip1 in A431 cells

In this paper, we provide evidence that both the mRNA and protein levels of the cyclin-dependent kinase (CDK) inhibitor p21WAF1/CDK-interacting protein 1 (Cip1) increase upon infection of A431 cells with Vaccinia virus (VACV). In addition, the VACV growth factor (VGF) seems to be required for the gene expression because infection carried out with the mutant virus VACV-VGF- revealed that this strain was unable to stimulate its transcription. Our findings are also consistent with the notion that the VGF-mediated change in p21WAF1/Cip1 expression is dependent on tyrosine kinase pathway(s) and is partially dependent on mitogen-activated protein kinase/extracellular-signal regulated kinase 1/2. We believe that these pathways are biologically significant because VACV replication and dissemination was drastically affected when the infection was carried out in the presence of the relevant pharmacological inhibitors.

Vasculite retiniana hemorragica multifocal aguda (Blumenkranz) / Acute multifocal hemorrhagic retinal vasculitis

Os autores descrevem 2 casos (4 olhos) com vasculite retiniana com características compatíveis com uma forma especial de vasculite idiopatia denominada vasculite retiniana heomorrágica multifocal aguda. Discutem a fisiopatia como também o exame de PCR (reaçäo em cadeia da polimerase) realizado em um caso.

HIV-1 subtyping by heteroduplex mobility assay in patients with bleeding disorders

O vírus da imunodeficiência humana do tipo 1 estabelece uma infecçäo presistente que na maioria dos casos evolui para a Síndrome de Imunodeficiência humana do tipo 1 tem sido geneticamente classificado em maior ou em outros grupos. Ogrupo maior é subdivido em nove subtipos baseados em evidências seqüênciais. A infecçäo pelo vírus da imunodeficiência humana do tipo 1 foi transmitida aos hemofílicos principalmente pelos concentrados de coagulaçäo no final da década de 70 e meados da de 80 e a síndrome da imunodeficiência adquirida tornou-se a principal causa de morbidade e morte entre estes pacientes. O Objetivo deste estudo foi o de determinar os subtipos do vírus da imunodeficiência humana em oito pacientes soropositivos com moléstias hemorrágicas de Belo Horizonte, Brasil, utilizando o ensaio de mobilidade heteroduplex, um método näo seqüêncial, que tem a propriedade de separar os portadores do vírus. Realizamos a reaçäo da cadeia de polimerase seguida pelo ensaio de mobilidade heteroduplex e obtivemos em todos os oito pacientes a confirmaçäo que os mesmos pertenciam ao subtipo B do vírus da imunodeficiência humana que é a mais prevalente nos Estados Unidos, Europa e Brasil. Os pacientes hemofílicos provavelmente foram infectados de concentrados provenientes da Europa, Estados Unidos, Säo Paulo e Rio de Janeiro (Brasil), utilizados em período anterior ao do conhecimento do vírus da imunodeficiência humana adquirida, e do uso de plasmas e concentrados näo testados, ou inativados, para o mesmo. O ensaio da mobilidade de heteroduplex e amplificaçäo do ácido desoxiribonucleico pela reaçäo de cadeia da polimerase provaram ser um modo rápido de subtipar o vírus da imunodeficiência humana adquirida em indivíduos infectados por transfusäo. O teste pode ser útil como rastreamento e detecçäo da origem e procedência do vírus da imunodeficiência adquirida.

PCR-based diagnosis of a case of herpetic whitlow in an AIDS patient

As infeccoes herpeticas sao complicacoes comuns em pacientes com AIDS. As manifestacoes clinicas podem ser incomuns e o tratamento antiviral e imperativo. Um metodo diagnostico rapido pode prevenir abordagens e tratamentos incorretos. A reacao em cadeia da polimerase (PCR) e um metodo rapido, sensivel e especifico para a amplificacao de DNA e para o diagnostico de doencas infecciosas, especialmente as de etiologia viral. Esta abordagem tem vantagens quando comparada com os metodos convencionais de diagnostico virologico. Recentemente nos relatamos um novo protocolo de PCR para o dignostico rapido de infeccoes herpeticas com supressao da etapa de extracao de DNA. Neste trabalho nos apresentamos um caso de paroniquea herpetica com diagnostico atraves de PCR especifico para Herpes Simplex tipo 1 usando o referido protocolo